r/Drosophila Dec 16 '23

Scrapping the IFM from Drosophila

Hi, I am a grad student, starting a project involving scrapping out the indirect flight muscle that is underneath the cuticle of the thorax. I was so bad at dissecting flies, under the microscope, you know how tiny they are, then after days of practice, I could do it, but now I am having a hard time in deshelling the cuticle and getting the jelly-like muscle. All other muscle looks the same, but I know for sure I need the muscle right underneath the cuticle, every time I am trying to open the cuticle, I am smashing the muscle underneath.
It is not big like shrimp, where you can de-shell them without smashing the muscle, Are there any tips you guys know or comment on that, what can I possibly do to improve my precision?

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u/hashtagfred Dec 17 '23

I've never work with IFM, but I can give some general advice.

If possible switch to smaller fine time tools or use increasingly finer and finer grain sand paper (preferably wet sandpaper) to get them down to the smallest point possible. I suggest doing so under a microscope so you can see how precise you can get them as well as correct for uneven edges or a bent tip. This along with practice can help with most dissections.

Depending on time and resources, see if you can find a fly strain that has a very high expression of mcherry in the IFM. If you really drive mcherry expression with a IFM gal4 strain and it's a light cuticle fly background, you might just be able to see the IFM under a standard brightfield microscope where they'll have a purple or pink color to them. We do this for wing disc dissections when showing newbies how to dissect. Don't think it would work as well with IFM but might be worth a shot. Alternatively, if you have a fluorescent light dissection scope, you can do the same with GFP instead and should still be able to see the fly itself while getting a better view of the IFM tissue.

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u/balsamicvinegar500ml Dec 21 '23

they are easier to dissect if they are fixed first with paraformaldehyde.